Molecular Biology Reports

BACKGROUND: Anemia is a negative factor in cancer, influencing the prognosis, quality of life and financial situation of cancer patients. Recent studies have shown that anemia in cancer is provoked by augmented erythrocyte removal. OBJECTIVE: In this study we sought to investigate the molecular bases for erythrocyte removal in cancer patients with anemia. In particular, we explored the levels of erythrocyte CD47, lactadherin, calreticulin and MCP1. METHODS: Thirty five anemic cancer patients (25 women, aged 66.4 +/-11.35 years old) and twelve healthy non-anemic controls (8 men, aged 61.1+/-9.98 years old) participated in our study. Red blood cells were isolated throug multiple centrifugations, and were lysed with the use of Triton-X 100. The levels of CD47, lactadherin, calreticulin and monocyte chemoattrractant protein 1 were determined by ELISA. RESULTS: Erythrocytes of anemic cancer patients display reduced CD47 (p<0.001), MCP1 (p=0.05), CD47 to lactadherin ratio (p<0.05), and increased lactadherin levels (p<0.01) in comparison to the healthy controls. DISCUSSION: Reduced CD47 along with increased lactadherin possibly drive erythrocyte removal in anemic cancer patients. Conclusions: The role of CD47 and increased lactadherin should be examined in the future as potential therapeutic targets and biomarkers for anemia diagnosis and iron dymsetabolism.

BackgroundReactive Oxygen Species (ROS) are the small, unstable and highly reactive species, having DNA oxidizing ability. Oxidation of the DNAs purine and pyrimidine bases can lead to single or double strands in this macromolecule. In this situation, the ATM molecule, a serine-threonine kinase, targets several proteins for phosphorylation, which causes the cell cycle to stop and the DNA damage repair begins. It has previously been proven that natural polyphenols have the cancer inhibiting properties due to their high efficacy and low side effects. Silibinin is the main herbal and medical ingredient in Milk Thistle (Silybum marianum) is a polyphenol flavonolignan, which has been widely considered as an antioxidant and anticancer agent. The purpose of the present study was to investigate the ATM gene expression and measurement of reactive oxygen species (ROS) in SKBR3 cell line, treated with Silibinin. Materials and MethodsAt first, the SKBR3 cell line was cultured in RPMI1640 culture medium and MTT assay was carried out to evaluate the Silibinin cytotoxicity. Flow Cytometry was carried out for cell cycle analysis, apoptotic induction, and ROS detection. While, Real Time PCR was used to evaluate the ATM gene expression in the Silibinin-treated and un-treated SKBR3 cells. ResultsPresent results have shown that 150 {micro}M Silibinin had the most significant cytotoxicity and apoptotic induction influence after the treatment period of 48 h. Flow cytometry data have shown that Silibinin induced considerable amount of apoptosis and caused cell cycle arrest at G1/S phase and induced production of ROS. Real-time PCR results have revealed that Silibinin increased the ATM expression in SKBR3 cell line. ConclusionSilibinin causes increased ATM gene expression by inducing ROS production, which initiates cell cycle arrest and apoptotic induction in SKBR3 cells line.

Background with objective: Detecting EGFR mutations is critical for treating lung adenocarcinoma with highly effective targeted therapies. However, standard genetic testing is expensive, complex, and often unavailable in resource-limited settings like Bangladesh. Because elevated serum CEA has been linked to these genetic alterations, it could serve as an accessible screening tool. This study aims to evaluate the association between serum CEA levels and EGFR mutation status to determine if routine CEA testing can reliably predict these mutations and guide treatment. Methodology: In this cross-sectional analytical study, we recruited 58 patients with histologically confirmed treatment naive lung adenocarcinoma. The presence of EGFR mutations in the ctDNA was determined via ARMS (Amplification Refractory Mutation System) PCR. Patient data was statistically analyzed to assess the diagnostic correlation between serum CEA levels and the presence of EGFR mutations. Result: The overall EGFR mutation rate was 43.1% with exon 19 deletion (48%) and exon 21 mutations (44%) were the predominant types. Median serum CEA levels were significantly higher in patients with EGFR mutations compared to wild-type cases (14.6 ng/ml vs 2.8 ng/ml, p<0.001). A multivariate analysis revealed a 14% increased likelihood of an EGFR mutation for 1 ng/ml rise in serum CEA. Furthermore, serum CEA showed strong diagnostic accuracy for ctDNA samples at a 6.39 ng/ml cut-off (AUC 0.82, sensitivity 68.0%, specificity 84.8%). Conclusion: Serum CEA is a valuable, cost-effective, and non-invasive biomarker demonstrating significantly higher levels and strong diagnostic accuracy in EGFR-mutated lung adenocarcinoma compared to wild-type cases.

Abstract Aim: Low chemotherapy response is a major risk factor for early mortality in cancer patients; it is one of the biggest challenges in cancer treatment. Main aim of this study is to identify chemotherapy non-responder, prognostic significance of pre-chemotherapy baseline variables in survival, distinguish most effective anti-cancer drug classes and formulation. Methods: In this multi-center retrospective cohort (n=2459) patients deceased with NSCLC and received anti-cancer drugs were included for analyses. To identify chemotherapy non-responder, patient population was divided into three sub-groups based on chemotherapy prescription frequency [1-15] as group-A, [16-30] as group-B, and [[&ge;]31] as group-C. Multivariate analysis was performed to identify risk of 1-year mortality in these groups. To prognose chemotherapy response in resected and unresected NSCLC patients, 0-7 days pre-chemotherapy white blood cell (WBC) count total five-ranges were compared as per overall survival in abnormal Vs normal WBC counts. Results: Post-stratification in group-A there were (n=1289) patients, in group-B (n=648) patients, and in group-C (n=522) patients. In group-A (n=301) patients 23% were found to have no new metastasis post-diagnosis significantly less p-value (0.004) compared to Group-B (n=125) 19.3%, and group-C (n=110) 19.2% patients p-value (0.008). Metastasis during chemotherapy was found significantly less in 20% patients of group-A, compared to (33%) in group-B, and (43%) in group-C p-value (<0.001). Post-chemotherapy initiation OS in group-A patients were significantly less 9 months (95% CI 9.3 - 9.6) compared to group-B 19 months (95% CI 17.7 - 20.2) and group-C 36.6 months (95% CI 34.6 - 38.5) patients p-value (<0.0001). Despite of low new metastasis and post chemo metastasis, group-A patients survived significantly less based on these outcomes group-A patients were considered as chemotherapy non-responder. Males and NSCLC stage III/IV patients were at higher risk; clinical benefits are corelated to surgery and radiotherapy for chemotherapy non-responder. Leukocytosis in both resected/unresected NSCLC group-A (13%) patients were found to be bad prognostic factor of survival in unresected group-B (5%) patients. Oral formulation of receptor tyrosine kinase inhibitors (RTKI) was effective in non-responders. Conclusion: Stratification of patient population based on chemotherapy prescriptions could be a useful method to find chemotherapy response in retrospective analysis. Patients with pre-chemotherapy leukocytosis should be closely monitored prior to selection of chemotherapy dose and formulation.

Objective To develop and validate a risk model for predicting postoperative bleeding in patients with thyroid cancer. Methods A total of 2800 consecutive patients diagnosed with thyroid cancer in the Department of Thyroid and Breast Surgery of the Affiliated Hospital of Xuzhou Medical University between January 2020 and December 2023 were retrospectively analyzed. Patients were categorized into two groups based on postoperative bleeding occurrence: bleeding and non-bleeding groups. Univariate and multivariate logistic regression analyses were utilized to screen independent risk factors. Meanwhile, risk prediction models were developed and nomogram . Subgroup analysis was performed to identify independent risk factors. The predictive effects of the models were assessed using the Hosmer-Lemeshow test and receiver operating characteristic (ROC) curves. Results Of the 2800 recruited patients, 50 had postoperative bleeding, with an incidence rate of 1.7%. Multivariate logistic regression analysis showed that age, hypertension, total thyroidectomy, tumor size [&ge;]4 cm, and operation time [&ge;]90 min were the risk factors for postoperative bleeding in thyroid cancer patients (P<0.05). A risk prediction model was established based on the above factors, and the area under the ROC curve was 0.881, with a sensitivity of 94.0%, a specificity of 67.3%, and an accuracy of 74.0%. Decision curve analysis revealed that the model had good predictive ability. Conclusions The constructed risk prediction model has good predictive power and can provide a reference for healthcare professionals to predict the risk of bleeding in patients after thyroid cancer surgery.

Understanding the genetic structure of keystone species within river networks is essential for effective conservation and management. While population differentiation of anadromous species often occurs between river systems, less research has been conducted on differentiation within rivers with smaller catchment areas. In this study, we investigated the population genetic structure of wild Atlantic salmon (Salmo salar) across the small-scale river Ronne [a] system in southernmost Sweden using Restriction-site Associated DNA sequencing (RADseq). Although the Admixture analysis did not detect clearly defined genetic clusters, significant pairwise FST values and DAPC revealed emerging population differentiation among the Ronne [a] tributaries. The observed patterns are consistent with a system characterized by connectivity, where genetic flow is present but can be reduced by behavioral and ecological factors such as spawning homing behavior and selective movements. These findings suggest that, despite overall connectivity, Atlantic salmon populations in the Ronne [a] catchment area may function as partially independent sub-populations. This highlights the importance of conservation and management strategies in fragmented river systems to consider population genetic structure to support resilient salmon populations under ongoing anthropogenic pressures.

Background Lung cancer mortality is rising in Libya, but access to molecular diagnostics for EGFR mutations--essential for guiding tyrosine kinase inhibitor therapy--remains severely limited. Selecting an appropriate testing platform requires balancing analytical performance against cost and infrastructure constraints. Methods We conducted a prospective comparative validation study using formalin-fixed paraffin-embedded (FFPE) tissue samples from Libyan non-small cell lung cancer (NSCLC) patients. Following stringent DNA quality control, samples were tested in parallel across four platforms: multiplex real-time PCR (MRT-PCR), reverse hybridization strip assay (RHSA), agarose gel electrophoresis (AGE), and immunohistochemistry (IHC). Performance was assessed by inter-method concordance, turnaround time, and cost per test. Results Of 30 initial samples, only six (20%) met quality thresholds (A260/A280 1.70-1.90; concentration [&ge;]10 ng/{micro}L), highlighting pre-analytical challenges. Three samples harbored EGFR exon 19 deletions. A critical discordance was identified: one sample tested negative by MRT-PCR (Ct {approx}38, {Delta}Ct=13) but positive by RHSA, AGE, and IHC, indicating a false-negative result from the reference method. IHC and RHSA offered the most favorable balance of cost (USD 40-75/test) and operational feasibility, while MRT-PCR (USD 150/test) required specialized infrastructure. Conclusions Relying solely on automated PCR may lead to under-diagnosis in low-cellularity or degraded FFPE samples. We recommend a hybrid algorithm: IHC as a cost-effective primary screen, followed by RHSA for confirmation. This approach optimizes resource allocation and improves diagnostic equity in Libya.

STRUCTURED ABSTRACT Purpose of Research: The generation of iPSC lines expressing 21, 56 and 79 glutamine repeats within the HTT protein and homozygous KO of HTT in the KOLF2.1J background as an additional disease series within the iPSC Neurodegenerative Disease Initiative (iNDI) collection. Major Findings: All iPSCs, even those expressing long repeats of 79Q or HTT KO, were capable of differentiating to striatal and cortical neurons, astrocytes and microglia using established protocols. General quality control stains and morphological analyses are described for each differentiation. A selected set of assays were carried out on differentiated cells; expanded repeat expressing astrocytes showed altered expression of astrocyte protein markers and morphological characteristics, and striatal neurons showed altered DARPP-32/CTIP2 colocalization. mRNAseq carried out for striatal neurons showed high similarities in gene expression changes between 79Q and KO lines compared to the unexpanded repeat. Conclusions: The KOLF2.1J isogenic CAG repeat series serves as a community resource to study HD mechanisms with the potential for direct comparison across other neurodegenerative diseases through the iNDI collection.

Objective: The APOE {varepsilon}4 genotype is the known most significant risk factor for sporadic Alzheimer's disease (AD), but the association between the blood levels of its encoded apolipoprotein E4 (ApoE4) and AD is poorly understood. Methods: We developed a chemiluminescent quantitative assay and a colloidal gold lateral chromatography qualitative assay to detect ApoE4 protein in plasma samples and compared the results with conventional genotyping. Results: In 185 samples, the plasma ApoE4 protein levels in non-{varepsilon}4 carriers ({varepsilon}2/{varepsilon}3 and {varepsilon}3/{varepsilon}3) were 0.07 +/- 0.23 and 0.05 +/- 0.17 g /mL, respectively; while the levels in {varepsilon}4 carriers ({varepsilon}2/{varepsilon}4, {varepsilon}3/{varepsilon}4, and {varepsilon}4/{varepsilon}4) were 4.58 +/- 1.96, 4.28 +/- 3.49 and 8.27 +/- 9.04 g/mL, respectively. Using a threshold of 1.1 g /mL, the quantitative method achieved a sensitivity of 100% and a specificity of 99.2%, while the qualitative method showed a concordance rate of 96.2% with genotyping. We also observed that among 133 non-{varepsilon}4 carriers, one had plasma ApoE4 protein levels higher than the threshold, which would require further investigation. Conclusion: Quantitative and qualitative detection of ApoE4 protein provides as rapid and accurate method for identifying APOE {varepsilon}4 genotype carriers, offering valuable insights for Alzheimer's disease risk assessment and early intervention.

Emerin (EMD), an inner nuclear membrane protein essential for nuclear architecture integrity, gene expression, cellular signaling, and chromatin stability, interacts with the LINC complex and participates in cytoskeleton-nucleoskeleton communication by binding to nuclear actin filaments. EMD is implicated in migration, invasion, and metastasis in some tumors, but its role in glioblastoma (GBM) remains unclear. This study evaluated the effects of EMD knockdown and overexpression in GBM cell lines following genotoxic treatment with cisplatin. In both wild-type p53 (U87-MG) and mutant p53 (U138-MG) GBM cells, EMD expression is high, and cisplatin treatment did not affect these protein levels. EMD knockdown in U87-MG cells significantly increased cisplatin IC50, viability, and proliferation. Conversely, stable overexpression of EMD in U87-MG cells led to reduced cisplatin IC50, viability, proliferation, and migration. EMD knockdown or overexpression did not affect any U138-MG phenotypes, with or without cisplatin treatment. Modulation of EMD levels causes morphological changes in stress fiber cytoskeleton, whereas overexpression of EMD in U87-MG cells promotes an increase and a decrease in nuclear and cytoplasmic actin levels, respectively. These biological responses of U87-MG cells overexpressing EMD were coincidentally associated with alterations in the levels of pH2AX(Ser139), p-p53(Ser15), p53, and p21Kip1 proteins after cisplatin exposure. In sum, modulation of EMD levels affects the viability, migration, and proliferation of wild-type p53 GBM cells treated with cisplatin, suggesting unknown roles in the DNA damage response and repair. This work highlights EMD as a potential regulator of GBM chemoresistance and a target for therapeutic intervention.

BackgroundHeterochromatic long arm of mouse Y chromosome harbors the multicopy species-specific sequences Ssty, Sly, Asty and Orly that are transcribed in testis and have known functions in male fertility. Of these Ssty and Sly encode proteins - yet all the transcripts are not translated. To investigate the roles of these Y-heterochromatic transcripts further, we analyzed them. MethodsMice with 2/3rd deletion of the Y-chromosome (XYRIIIqdel) and its wild type (XYRIII) were used in this study. Bioinformatic approaches, small RNA northern blots, Electrophoretic Mobility Shift Assays, Luciferase reporter assays, dPCR analysis, RT-qPCR assays and western blotting techniques were used to identify piRNAs that regulate autosomal genes. ResultsWe demonstrate that the multicopy gene families from mouse Y-long arm generate piRNAs predominantly in testis. We observed sequences homologous to these piRNAs in the UTRs of a few autosomal genes, which are differentially expressed in the sperms of XYRIIIqdel mice. Furthermore, the Endogenous Retrovirus Element (ERV) LTR, found in the Orly1 transcript identified piRNAs in the database, showed homology to UTRs and associated genomic regions of a few autosomal genes. Orly1 showed a reduction in genomic copy number by digital PCR in XYRIIIqdel mice. One of the four autosomal genes containing the ERV segment in their UTRs, showed a differential testicular protein expression in the mutant mice. ConclusionsThus, we further elucidate that different classes of repeats from Y-chromosome regulate autosomal gene expression via piRNAs. Besides, this study also identified novel roles for a Y-derived ERV in autosomal gene regulation in testis.

The AMY2B gene encodes pancreatic amylase, a critical enzyme for starch digestion. While previous studies have examined AMY2B copy number variation (CNV) in domestic and some wild animals, less is known about wild carnivores inhabiting regions with limited anthropogenic starch exposure. We analyzed blood samples for serum amylase activity and copy number variation in AMY2B gene from 8 wolves (Canis lupus), 11 brown bears (Ursus arctos), and 3 red foxes (Vulpes vulpes) from Bosnia and Herzegovina. AMY2B gene copy number was assessed using droplet digital PCR (ddPCR), and serum amylase activity and glucose levels were quantified. Although the number of fox samples was limited, foxes and wolves consistently harbored two copies of AMY2B, while brown bears exhibited higher CNV (3.67-8.40, mean 5.88). Serum amylase activity was highest in foxes, moderate in wolves, and variable but lower in bears. Despite differences in AMY2B copy number and serum amylase activity, circulating glucose concentrations did not differ significantly among species. Our findings suggest that variation in AMY2B copy number among wild carnivores may be associated with species-specific evolutionary histories and dietary adaptations, providing insight into genomic mechanisms underlying carbohydrate utilization in natural populations.

Stem cell therapy represents a promising strategy for the replacement and functional restoration of damaged neural tissue in neurodegenerative conditions. Human dental pulp stem cells (hDPSCs) have emerged as potential candidates for neuroregeneration due to their ease of isolation, neural crest origin, neurotrophic and anti-inflammatory capacity, and demonstrated ability to differentiate in vitro into neuronal-like cells exhibiting electrophysiological activity. Although the immunomodulatory and neuroprotective properties of hDPSCs have been reported in multiple models of brain disease, their capacity to functionally integrate into host neuronal circuits remain poorly understood. In this study, we have grafted green fluorescent protein (GFP)-transduced, neural preconditioned hDPSCs into the CA1 region of the hippocampus of C57BL/6J mice. One month after transplantation, GFP+-hDPSCs survived in the brains of non-immunosuppressed mice and remained localized within the grafted area. Notably, the transplanted cells underwent in situ differentiation and exhibited a neuroblast-like phenotype, characterized by positive doublecortin expression and immature neuronal-like electrophysiological properties, like high membrane input resistance, low capacitance, and the ability to generate single action potentials after stimulation. Together, these findings provide the first evidence that hDPSCs can survive and integrate into the hippocampal network of the mouse brain at one-month post graft, supporting their potential use for future therapeutic applications in acute brain lesions and neurodegenerative disorders.

It has been previously shown that chronic social defeat stress caused by paired agonistic interactions between male mice is accompanied by the development of depression-like state and immune deficiency. The aim of this study was to investigate changes in the expression of C1qtnf superfamily genes (encoding the complement component related with tumor necrosis factor) in the hypothalamus, thymus and lungs against the background of the Lewis lung adenocarcinoma growth. In the experiments, on the 5th day of social stress, male mice were injected with tumor cells into the tail vein. Chronic social stress continued for the next two weeks. The transcriptomes of the hypothalamus, thymus and lungs of mice were sequenced at the Genoanalytica Collective Center (http://genoanalytica.ru/, Moscow). Changes in the expression of the C1qtnf genes in the tissues of stressed mice were studied compared with the control and mice that were additionally injected with tumor cells. Overall, significant correlations were found between expression of most genes in each tissue of the experimental groups. In the hypothalamus of stressed animals, when tumor cells were introduced, an increase in the expression of the genes C1qtnf1, C1qtnf2, C1qtnf3, C1qtnf6 and C1qtnf7 was observed compared to controls. In the thymus of these animals, tumor cell injection increased expression of the C1qtnf1, C1qtnf5, and C1qtnf6 genes. In the lung of tumor-injected stressed mice, expression of the C1qtnf1, C1qtnf2, C1qtnf7, and C1qtnf9 genes was decreased relative to controls and non-tumor-injected depressed mice, reaching near-zero levels in some mice. Analysis of C1qtnf superfamily gene expression in the all tissues revealed negative correlations between the expression of the C1qtnf1, C1qtnf2, and C1qtnf7 genes in the hypothalamus and lungs indicating synchronization of processes against the background of social stress and Levis lung adenocarcinoma.

IntroductionThe proteasome is a critical cellular degradative machinery impaired in late-stage Alzheimers disease (AD). However, the status and activity of the proteasome in early-stage sporadic AD (sAD) is unknown. MethodsA cellular model of human early-stage sAD was generated from sAD patient iPSC-derived cortical neurons by dual-SMAD inhibition. The iPSCs, neuroprogenitors, and cortical neurons were validated by the expressions of key markers. The level of total intraneuronal A{beta} was measured by ELISA. Composition and native proteolytic activities of the proteasome in control and sAD cortical neurons were measured using complementary fluorogenic probes. ResultsControl and sAD patients iPSCs expressed pluripotent markers OCT4, NANOG, and SSEA4 which induced into neuroprogenitors expressing NESTIN and PAX6. The neuroprogenitors terminally differentiated into cortical neurons expressing neuronal markers MAP2 and TUJ1, and cortical layer marker TBR1. The level of intraneuronal A{beta} in the sAD cortical neurons was significantly higher compared to control. Control and sAD cortical neurons expressed native 30S, 26S, and 20S proteasome assemblies with the sAD cortical neurons displaying higher 20S assemblies. Increased active 20S assemblies was associated with higher {beta}1, {beta}2, and {beta}5 proteolytic sites activities. DiscussionThe significant elevation in the proteolytic activities of the {beta}1, {beta}2, and {beta}5 subunits of 20S proteasome in sAD cortical neurons suggests that this may be a possible compensatory response to elevated intraneuronal A{beta}. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=120 SRC="FIGDIR/small/734021v1_ufig1.gif" ALT="Figure 1"> View larger version (30K): org.highwire.dtl.DTLVardef@1d8c382org.highwire.dtl.DTLVardef@b92e8org.highwire.dtl.DTLVardef@1d9c699org.highwire.dtl.DTLVardef@7d826d_HPS_FORMAT_FIGEXP M_FIG C_FIG

Background: This study aimed to elucidate B cell subset pathology in COPD, a poorly characterized area, with a focus on its similarities to and differences from classical autoimmune disorders. Methods: Single-cell RNA-sequencing (scRNA-seq) data from COPD and autoimmune diseases were obtained from the Gene Expression Omnibus (GEO) for comparative analyses of B cell subsets and functions via differentially expressed genes (DEGs), KEGG, protein-protein interaction (PPI), and cell-cell communication analyses. Serum IgG4 was measured by ELISA and correlated with clinical parameters. Peripheral blood B cells were sorted by flow cytometry for single-cell B cell receptor (BCR) sequencing. A v-Abl-Bcl2 pro-B cell line was stimulated with cigarette smoke extract (CSE) to assess abnormal development in vitro. Results: In lung tissue, IgG4 plasma cells were enriched and expressed BCR activation and inflammatory genes and TNF-NF-kB-MAPK pathways. Serum IgG4 concentrations correlated negatively with pre- and post-bronchodilator FEV1-FVC. B cells interacted with monocytes, macrophages, fibroblasts, and endothelial cells via IL-1B-IL-6, integrin, and chemokine signaling, contributing to chronic inflammation and remodeling. In peripheral blood, transitional T1 B cells were increased, accompanied by lambda-chain enrichment and increased IGLV1-47 usage, as well as enrichment of autoimmune pathways. In the bone marrow, the numbers of pre-B I cells were increased while those of small pre-B III cells were reduced, with altered expression of BCR development genes. CSE stimulation of the pro-B cell line reduced lambda expression in a concentration-dependent manner. Conclusions: The autoimmune abnormalities in COPD appear more restricted, although IgG4 antibody generation may contribute to immune-mediated lung damage.

While recent therapeutic advances have extended the survival of patients with non-small cell lung cancer (NSCLC), overcoming metastatic progression in the CNS remains a significant challenge. Some patients with NSCLC may require concurrent management of CNS and extracranial metastases, while others develop isolated brain metastasis or leptomeningeal disease. These heterogenous clinical outcomes are difficult to predict and diagnose for early intervention with current surveillance modalities. Herein, we comprehensively analyzed gene mutations, copy number variations, and DNA methylation of NSCLC brain metastasis tissue collected at the time of craniotomy, combined with ctDNA sequencing of paired plasma and CSF liquid biopsies. We confirmed a high concordance between the molecular features of brain metastasis tissue with ctDNA from CSF which were largely distinct from ctDNA alterations in paired plasma samples. Plasma ctDNA tumor fraction and ctDNA hypermethylation were most significantly associated with extracranial metastasis and overall survival. Alternatively, we identified specific hypermethylated DNA loci in brain metastasis tissue and CSF ctDNA as significant correlates of brain metastasis progression and risk of leptomeningeal disease. Our findings support the utility of integrating ctDNA testing from CSF and plasma, while revealing distinct epigenetic features and biomarkers of brain metastasis or leptomeningeal disease.

Background Human T-cell lymphotropic virus - 1 (HTLV-1) is the causative agent of Adult T-cell Leukaemia/Lymphoma (ATLL), a malignancy of CD4+ cells, and HTLV-1-associated Myelopathy/Tropical Spastic Paraparesis (HAM/TSP), a demyelinating disease. Globally, 10-20 million people are infected, though most remain asymptomatic and about 5% progress to severe disease. Transmission occurs mainly through breastfeeding, sexual contact, contaminated needles, and blood transfusion. In Ghana, evidence on the role of HTLV-1 in haematological malignancies remains scarce. Methods This was a cross-sectional study involving 200 patients with haematological malignancies (Acute Lymphoblastic Leukaemia - 4, Acute Myeloid Leukaemia - 6, Chronic Lymphocytic Leukaemia - 27, Chronic Myeloid Leukaemia - 63, Hodgkin Lymphoma - 21, Multiple Myeloma - 31, Myelodysplasia - 6, Myeloproliferative Neoplasm - 11) at the Haematology Day Care of the Korle-Bu Teaching Hospital. After informed consent was obtained, sera from study participants were tested for anti-HTLV-1 using MP Diagnostics GmbH ELISA immunoassay. Data were analysed using R software version 4.0.2 and SPSS version 31.0.0. Results The study population had a mean age of 49.1{+/-}17.7 years, with majority being females (n=109, 54.5%). Of the 200 samples, 16 (8.0%) were seropositive for HTLV-1, and these were detected in 4 males and 12 females. No statistically significant association was found between HTLV-1 infection and haematological malignancy (exact p = 0.061), sex (p=0.061), and history of blood transfusion (exact p= 1.000). Conclusion The findings show the seroprevalence of HTLV-1 of 8.0% among patients with haematological malignancies. Although there was no probable association between HTLV-1 and haematological malignancies, screening for HTLV-1 in patients with haematological malignancies may help to unravel the exact contribution in these conditions.

Anoxia is a major stress for most vertebrates and frequently accompanies harsh winter conditions, particularly in species that spend much of the season frozen solid. North American freeze-tolerant wood frogs (Rana sylvatica) can survive several months without oxygen and endure whole-body freezing for up to eight months of the year, with [~]70% of total body water frozen as extracellular ice, yet revive when temperatures rise in spring. Survival depends on multiple adaptations, including tolerance of prolonged oxygen deprivation while frozen, when breathing and circulation are halted. A key strategy involves hepatic glycogen mobilization, producing large amounts of glucose that are distributed to tissues where it functions both as a cryoprotectant and as a substrate for anaerobic ATP production. The present study examines the role of histone lysine methylation and demethylation in regulating liver proteins under anoxic conditions. Relative protein expression of seven histone methyltransferases (ASH2L-S, ASH2L-L, RBBP5, SETD8, SMYD2, ESET, SETD1), six lysine demethylases (KDM1A, KDM3B, KDM4A, KDM4B, KDM5A, KDM5C), and eight histone marks (H3K4me1, H3K4me2, H3K9me3, H3K27me3, H3K36me3, H3K79me3, H4K20me1, H4K20me3) were evaluated in wood frog liver under control, 4-hour, and 24-hour anoxia exposures. The data indicate that histone lysine methylation and demethylation contribute significantly to transcriptional regulation under anoxia. Specifically, H3K4, H3K36, and H3K79 methylation were associated with transcriptional activation, whereas H3K9, H3K27, and H4K20 methylation correlated with transcriptional repression. These findings highlight the dynamic role of epigenetic regulation in supporting hypometabolism and stress adaptation in freeze-tolerant wood frogs.

Congenital disorders of glycosylation and deglycosylation are rare, serious, and lethal disorders afflicting humans. CDGs and CDDGs result in loss of function enzymes which fail to build or break down oligosaccharides on proteins. This can produce protein aggregates and, in turn, reactive oxygen species that harm the cell eventually leading to autophagy and apoptosis. Because sperm contain high concentrations of polyunsaturated fatty acids, they are especially sensitive to these effects, which is understood as one of the leading factors in human male infertility. Sperm are developed in zebrafish similarly to humans and are useful models to examine human reproductive health, as well as genetic disorders. The combination of these advantages makes the analysis of sperm from zebrafish with heterozygous ALG1 or DPAGT1 CDGs or the NGLY1 CDDG suitable. Analysis of sperm concentration, motility, status, viability, and hypoosmotic swelling demonstrated the effects of these disorders on sperm quality. Results showed a significant decrease in sperm concentration, motility, and hypoosmotic swelling for all mutant zebrafish compared to the wild type. This suggests that CDGs and CDDGs influence the amount of sperm produced, the percentage of sperm cells that are mobile, and the integrity of the plasma membrane.